Supplementary Materials Table S1 Primer sequences used in PCR analysis. mice. Data are expressed as means??SEM of 8 mice per group. *test (A) or TSPAN9 two\way ANOVA (C and D) was used. Physique S4 Metformin does not switch blood pressure or heart rate in either wild type or AMPK2?/? mice. Systolic blood pressure (SBP) of (A) wild type mice and (B) AMPK2?/? mice after 7?days of AngII infusion. (C) Diastolic blood pressure (DBP) of wild type mice and (D) AMPK2?/? mice after 7?days of AngII infusion. (E) Heart rate of wild type mice after 7?days of AngII infusion. (F) Heart rate of AMPK2?/? mice after 7?days of AngII infusion. test was used (C and D). Welch’s ANOVA with Games\Howell test was utilized for the other panels. Physique S5 Metformin has no effect on fasting blood glucose levels in wild type or AMPK2?/?mice. (A) Fasting blood glucose levels of wild type mice after 7?days of AngII infusion. (B) Fasting blood glucose levels of AMPK2?/?mice after 7?days of AngII infusion. Games\Howell test was used. Physique S7 AMPK2 knockout exacerbates AngII\induced HNF4 expression, TGF1 expression and cardiac fibrosis. (A) Left panel: western blot analysis of HNF4 expression in the heart. The right panel shows quantification from the HNF4 proteins amounts. (B) Quantitative true\period PCR evaluation of TGF1 mRNA appearance in center lysates. (C) The TGF1 proteins level was driven via elisa. (D) Still left -panel: representative micrographs of Sirius crimson\stained heart sections; the red area represents collagen. Bars =500?m. Ideal panel:Quantification of the fibrotic area is indicated as the CHIR-99021 tyrosianse inhibitor percentage of the total cardiac area. (E) Collagen I (remaining panel) and collagen III (ideal panel) mRNA manifestation was measured via actual\time PCR analysis. (F) Left panel: representative pulsed wave Doppler (PWD) images across the mitral circulation and cells Doppler (TD) images of the mitral valve ring within the 7th day time of AngII infusion in crazy type mice. Right panel: E/E. The E wave and E wave are indicated by arrows. Data are indicated as means??SEM of 8 mice per group. *test was used (B and C). Welch’s ANOVA with Games\Howell test was utilized for the additional panels. Number S8 Metformin did not improve cardiac diastolic function upon AngII exposure in AMPK2?/? mice. (A) Remaining panel: Representative PWD images showing the mitral circulation and TD images of the mitral valve ring within the 7th day time of AngII infusion in AMPK2?/? mice. Right panel: E/E. (B) Representative echocardiograms within the 7th day time of AngII or saline infusion in AMPK2?/? mice. (C) EF% within the 7th day time of AngII infusion in AMPK2?/? mice. (D) Remaining ventricular shortening portion (FS%) within the 7th day time of AngII infusion in AMPK2?/?mice. CHIR-99021 tyrosianse inhibitor Data are indicated as means??SEM CHIR-99021 tyrosianse inhibitor of 8 mice per group. *Games\Howell test (A) or two\way ANOVA (C and D) was used. BPH-175-1217-s001.pdf (1.4M) GUID:?56718445-5035-4261-A2D5-3571C9AD5F67 Abstract Background and Purpose Metformin, a small molecule, antihyperglycaemic agent, is a well\known activator of AMP\activated protein kinase (AMPK) and protects against cardiac fibrosis. CHIR-99021 tyrosianse inhibitor However, the underlying mechanisms remain elusive. TGF1 is definitely a key cytokine mediating cardiac fibrosis. Here, we investigated the effects of metformin on TGF1 production induced by angiotensin II (AngII) and the underlying mechanisms. Experimental Approach Wild\type and AMPK2?/? C57BL/6 mice were injected s.c. with metformin or saline and infused with AngII (3?mgkg?1day?1) for 7?days. Adult mouse cardiac fibroblasts (CFs) were isolated for experiments. Key Results In CFs, metformin inhibited AngII\induced TGF1 manifestation via AMPK activation. Analysis using.